PMID: 9182808Jun 1, 1997Paper

Cloning of human keratolinin cDNA: keratolinin is identical with a cysteine proteinase inhibitor, cystatin A, and is regulated by Ca2+, TPA, and cAMP

The Journal of Investigative Dermatology
H TakahashiHajime Iizuka

Abstract

Keratolinin has been described as one of the precursor proteins of cornified cell envelope of keratinocytes. Using rabbit polyclonal anti-human keratolinin antibody, we isolated a cDNA clone of human keratolinin gene from a human Agt11 cDNA expression library that was constructed by random priming from poly(A)+RNA extracted from cultured normal human keratinocytes. Screening by rabbit anti-human keratolinin antibody detected one positive clone (HKL-1). The recombinant 12.5-kDa protein constructed from the clone reacted specifically with the anti-human keratolinin antibody. DNA sequence analysis revealed that HKL-1 clone was 448 bp long, and its putative amino acid sequence was identical with that of a human cysteine proteinase inhibitor, cystatin A. Western blot analysis showed that the commercially available recombinant cystatin A also reacted specifically with the anti-human keratolinin antibody. Northern blot analysis indicated that HKL-1 clone hybridizes with mRNA of about 0.5 kb, consistent with the size of the HKL-1 clone. The keratolinin mRNA was highly expressed in cultured human keratinocytes in high Ca2+ (1 mM); in low Ca2+ (0.05 mM), the keratolinin mRNA expression was significantly lower. Using SV40-transformed huma...Continue Reading

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Citations

Mar 28, 1998·Experimental Dermatology·A Ishida-Yamamoto, H Iizuka
Nov 13, 1998·The Journal of Biological Chemistry·M F ChampliaudP F Olson
Jan 2, 2001·Critical Reviews in Oral Biology and Medicine : an Official Publication of the American Association of Oral Biologists·R B Presland, B A Dale
Nov 30, 2000·Biochemical and Biophysical Research Communications·H TakahashiH Iizuka
Mar 20, 1998·The Journal of Investigative Dermatology·H TakahashiH Iizuka
Feb 1, 2005·European Journal of Cell Biology·Patrick L J M Zeeuwen

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